| Hibernation is a way of survival for animals to cope with extreme harsh environment.During hibernation,the animal's body temperature,heart rate,respiration rate and most physiological activities are inhibited.At present,the hibernation regulation mechanism of Tamias sibiricus is not very clear,and the study on hibernation proteome of T.sibiricus is rarely reported.In this study,TMT labeling technology and quantitative proteomics technology were used to analyze and compare the changes of protein expression in the heart and liver of T.sibiricus during hibernation and active arousing phase,so as to further explore the regulatory mechanism of hibernation and how to deal with the body injury caused by ischemia and reperfusion.The main findings are as follows:1.A total of 40 significantly upregulated differential proteins(higher expression levels during hibernation)and 50 significantly down-regulated differential proteins(lower expression levels during hibernation)were identified in the chipmunk liver tissues.During hibernation period,the expression levels of enzymes related to carbohydrate metabolism(such as GSK3 and GP)remained unchanged or decreased,while the expression levels of lipid metabolism-related proteins(ACAA and HMG-Co A)increased significantly,indicating that the main energy supply mode of hibernating T.sibiricus tetrus was transformed from glucose metabolism to lipid decomposition,which was the same as that of other small hibernating animals.2.A total of 24 significantly up-regulated proteins and 46 significantly down-regulated proteins were identified in the heart tissues of the chipmunk.Most of the enzymes related to glucose metabolism remained unchanged or slightly decreased,while the expression levels of enzymes related to lipid metabolism did not change significantly during and after hibernation,which was not consistent with the changes in liver energy metabolism.3.Oxidative stress caused by ischemia-reperfusion is a challenge that hibernating animals must deal with.In the hibernating chipmunks,the contents of heat shock proteins and related proteins(HSP and PKC,etc.)were significantly increased in liver and heart,which are supposed to remove excess oxygen free radicals,and they may be the main substances to deal with ischemia-reperfusion injury in chipmunks.The increased expression of Anx A1 in the liver is also one of the ways to deal with excess oxygen free radicals,and it can also inhibit many inflammatory pathways and protect the body's health.The expression levels of c TN and GPBB in the heart tissues were normal,confirming that the heart of chipmunk was not damaged during hibernation.4.The diversity and structural composition of gut microorganism were analyzed by 16 S amplicon sequencing before and during hibernation.There was no significant difference in ? diversity index(Chao1,Simpson and Shannon),but there was significant difference in ? diversity.The dominant bacteria phyla were Bacteroidetes before and during hibernation,while Proteobacteria had a larger proportion during hibernation.Lefse's results showed that the Bacteroidetes with higher abundance in the hibernating group were concentrated.COG functional annotations mainly focus on carbohydrate transport and metabolism,transcription,and amino acid transport and metabolism.KEGG cluster analysis showed that the number of down-regulation in hibernation group was higher than that in active group,and only the sensory system,metabolic disease and excretory system were significantly up-regulated.Based on quantitative proteomic techniques,this study for the first time elucidates the proteomic characteristics and changes of heart and liver and gut microbiota before during hibernation of T.sibiricus,and reveals the proteomic and macrobiotic basis of hibernation of Siberian chipmunks,which provides important reference for us to understand the regulation mechanism of hibernation in small mammals,and also has reference significance for the treatment of many diseases. |
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